Littman B H, Muchmore A V, Steinberg A D, Greene W C
J Clin Invest. 1983 Dec;72(6):1987-94. doi: 10.1172/JCI111163.
Autoantibody-secreting hybridomas were produced by somatic cell fusion of B lymphocytes from a patient with systemic lupus erythematosus with two different human myeloma lines. Selection of hybrids formed from one of these cell lines was performed by using aminopterine-containing culture medium as this cell line was deficient in hypoxanthine-guanine-phosphoribosyl transferase (HGPRT). The second myeloma line was not HGPRT-deficient but instead was treated with diethylpyrocarbonate, which assured death of unfused myeloma cells. This novel technique has wide applicability. Hybridomas were found to secrete antibodies to native DNA and to extractable nuclear antigen. The binding specificities of one IgM anti-DNA antibody was characterized and found to be specific for double-stranded DNA and had particular binding affinity for poly(dG) . poly(dC).
通过将系统性红斑狼疮患者的B淋巴细胞与两种不同的人骨髓瘤细胞系进行体细胞融合,产生了分泌自身抗体的杂交瘤。由于其中一种细胞系缺乏次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(HGPRT),因此使用含氨基蝶呤的培养基来选择由该细胞系形成的杂交瘤。第二种骨髓瘤细胞系不缺乏HGPRT,而是用焦碳酸二乙酯处理,这确保了未融合的骨髓瘤细胞死亡。这种新技术具有广泛的适用性。发现杂交瘤分泌针对天然DNA和可提取核抗原的抗体。对一种IgM抗DNA抗体的结合特异性进行了表征,发现它对双链DNA具有特异性,并且对聚(dG).聚(dC)具有特殊的结合亲和力。